Sell C, Dumenil G, Deveaud C, Miura M, Coppola D, DeAngelis T, Rubin R, Efstratiadis A, Baserga R
Jefferson Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
Mol Cell Biol. 1994 Jun;14(6):3604-12. doi: 10.1128/mcb.14.6.3604-3612.1994.
Fibroblast cell lines, designated R- and W cells, were generated, respectively, from mouse embryos homozygous for a targeted disruption of the Igf1r gene, encoding the type 1 insulin-like growth factor receptor, and from their wild-type littermates. W cells grow normally in serum-free medium supplemented with various combinations of purified growth factors, while pre- and postcrisis R- cells cannot grow, as they are arrested before entering the S phase. R- cells are able to grow in 10% serum, albeit more slowly than W cells, and with all phases of the cell cycle being elongated. An activated Ha-ras expressed from a stably transfected plasmid is unable to overcome the inability of R- cells to grow in serum-free medium supplemented with purified clones. Nevertheless, even in the presence of serum, R- cells stably transfected with Ha-ras, alone or in combination with simian virus 40 large T antigen, fail to form colonies in soft agar. Reintroduction into R- cells (or their derivatives) of a plasmid expressing the human insulin-like growth factor I receptor RNA and protein restores their ability to grow with purified growth factors or in soft agar. The signaling pathways participating in cell growth and transformation are discussed on the basis of these results.
分别从小鼠胚胎中生成了成纤维细胞系，命名为R细胞和W细胞。这些胚胎对于编码1型胰岛素样生长因子受体（Igf1r）的基因的靶向破坏是纯合的，以及来自它们的野生型同窝仔。W细胞在补充了各种纯化生长因子组合的无血清培养基中正常生长，而危机前和危机后的R细胞不能生长，因为它们在进入S期之前就停滞了。R细胞能够在10%血清中生长，尽管比W细胞生长得慢，并且细胞周期的所有阶段都被延长。从稳定转染的质粒表达的活化Ha-ras不能克服R细胞在补充了纯化克隆的无血清培养基中生长的无能。然而，即使在有血清的情况下，单独或与猿猴病毒40大T抗原一起稳定转染Ha-ras的R细胞也不能在软琼脂中形成集落。将表达人胰岛素样生长因子I受体RNA和蛋白质的质粒重新导入R细胞（或其衍生物）可恢复它们在纯化生长因子存在下生长或在软琼脂中生长的能力。基于这些结果讨论了参与细胞生长和转化的信号通路。
